Peptide antigens: a practical approach
Peptide antigens and anti-peptide antibodies are widely used in biochemistry and molecular biology for the measurement, location, and purification of specific oligopeptides. More recently the application of these reagents has expanded, for example in the identification and mapping of the binding sit...
Gespeichert in:
| Format: | Buch |
|---|---|
| Sprache: | English |
| Veröffentlicht: |
Oxford u.a.
IRL Press
1994
|
| Ausgabe: | 1. ed. |
| Schriftenreihe: | The practical approach series
144 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Zusammenfassung: | Peptide antigens and anti-peptide antibodies are widely used in biochemistry and molecular biology for the measurement, location, and purification of specific oligopeptides. More recently the application of these reagents has expanded, for example in the identification and mapping of the binding sites of antibodies and T-cell receptors and in the identification and characterization of proteins which are known only by their primary structure. This volume provides practical guidance to the major techniques used in the exploitation of peptide antigens and anti-peptide antibodies. The chapters give detailed protocols for the prediction of epitopes, peptides synthesis, the preparation of peptide immunogens, immunoaffinity chromatography, immunoassays, and the mapping of epitopes using both synthetic peptides and phage display systems Peptide Antigens: A Practical Approach covers all practical aspects of this important and growing subject. It is a unique compendium of methods for workers in biochemistry, molecular biology, and immunology who need to use this technology in their research |
| Beschreibung: | XIX, 252 S. graph. Darst. |
| ISBN: | 0199634521 0199634513 |
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| 520 | 3 | |a Peptide antigens and anti-peptide antibodies are widely used in biochemistry and molecular biology for the measurement, location, and purification of specific oligopeptides. More recently the application of these reagents has expanded, for example in the identification and mapping of the binding sites of antibodies and T-cell receptors and in the identification and characterization of proteins which are known only by their primary structure. This volume provides practical guidance to the major techniques used in the exploitation of peptide antigens and anti-peptide antibodies. The chapters give detailed protocols for the prediction of epitopes, peptides synthesis, the preparation of peptide immunogens, immunoaffinity chromatography, immunoassays, and the mapping of epitopes using both synthetic peptides and phage display systems | |
| 520 | |a Peptide Antigens: A Practical Approach covers all practical aspects of this important and growing subject. It is a unique compendium of methods for workers in biochemistry, molecular biology, and immunology who need to use this technology in their research | ||
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Datensatz im Suchindex
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| adam_text | Peptide Antigens
A Practical Approach
Edited by
GBRIANWISDOM
Division ofB iochemis t ry , School ofB io logy and Biochemistry
The Queen s Univers ity, Medica l Bio logy Cent re , Bel fas t , UK
at
OXFORD UNIVERSITY PRESS
Oxford New York Tokyo
Contents
List of contributors xv
List of abbreviations xvii
1 Peptide antigens and anti-peptide antibodies i
G Brian Wisdom
1 Anti-peptide antibodies and their applications 1
2 Peptide antigens and their applications 3
3 Developments 4
References 5
2 Epitope predictions from the primary
structure of proteins i
Jean-Luc Pellequer, Eric Westhof, and Marc H V Van Regenmortel
1 Introduction 7
Construction of prediction profiles 9
Comparison of prediction profiles 10
2 Common methods for predicting B-cell epitopes 10
3 Methods of predicting T-cell epitopes 15
4 Location of B-cell epitopes in proteins of known tertiary
structure 16
5 Comparison between prediction efficacies of different
scales 16
6 Epitope prediction based on turn prediction 19
7 Correlation between turn prediction am 1 antigenicity 20
8 Conclusions 22
References 23
3 Solid-phase peptide synthesis
Brian Walker
1 Introduction
Conten t s
2 The Merrifield (Boc) methodologv
General strategy and choice of functionalized resin
Chemical procedures
Cleavage of peptide from the solid support and removal of
side-chain protecting groups
3 The Fmoc methodology
General strategy
Chemical procedures
Protection of side-chain functional groups
Activation of amino acids and coupling procedures
Monitoring the extent of the acylation reactions
Cleavage of peptide from resin and side-chain deprotection
4 Methods of disulphide bridge formation
5 Purification of peptides
Characterization of purified peptide
i
6 Synthesis of phosphorylated peptides
References
4 Immunization with pepti de-carrier
complexes: traditional and multiple-antigen
peptide systems
James P Tarn
1 Introduction
2 Traditional and MAP approach? 3
Selection of peptides
B- and T-cell epitopes
3 Traditional approaches
Types of protein carrier
Conjugation and polymerization methods
Do s and don ts
4 The MAP approach
The MAP concept
The MAP strategy
The importance of peptide orientation
The effect of the number of branches
Characteristics of immune responses to MAPs
MAP applications to vaccines
5 Immunization procedures
Production of antibodies
Preparation and storage of serum and ascitic fluid
References
x
7(i
Contents
5 Preparative immunoaffinity techniques 117
Michael R Price and Kami Beyzavi
1 Introduction 117
General principles of immunoaffinity chromatography 117
2 Choice of gels and conjugation procedures 118
Theoretical requirements 118
Natural matrices 118
Synthetic matrices 119
Activation chemistry 120
Small ligands and spacer options 121
Protein A bead-linked antibodies 121
3 Chromatographic procedures 122
Equipment 122
Sample preparation and application 123
Column washing 124
Column elution 125
Quality control testing 125
4 Antibodies 126
Sources of antibodies 126
Storage and stability 126
5 Antibody purification 127
IgG monoclonal antibodies 127
IgG from serum 127
IgM monoclonal antibodies 129
Antigen affinity chromatography 129
Epitope affinity chromatography 129
Affinity purification with biotinylated peptides 131
6 Antigen purification 131
Preliminary considerations 131
Storage and stability 132
Purification of soluble antigens 132
Purification of membrane-associated antigens 133
7 Conclusions 135
Acknowledgements 136
References 136
6 Immunoassays of proteins and anti-peptide
antibodies 139
Nigel P Groome
1 Introduction 139
Why use anti-peptide antibodies? 139
xi
Contents
2 Immunoassays for the detection of antibodies 141
Detection of antibodies to parent proteins which react with
immobilized peptides 141
Detection of antibodies to peptides which react with immobilized
peptides 146
Detection of antibodies to peptides which react with immobilized
parent proteins 147
Selection of monoclonal anti-peptide antibodies with the highest
functional affinity for the parent protein 149
Peptide-carrier conjugates in immunoassay 150
Biotinylated peptides in immunoassay 151
Applications of branched peptides 153
The importance of peptide purity 153
3 Immunoassays for the parent protein using anti-peptide
antibodies 153
Development of immunoassays for molecules unavailable for
antibody production 153
Purification of antibodies for use in immunoassays 154
Competitive assays 156
Two-site (sandwich) assays 159
4 Immunoblotting 173
Electroblotting 173
5 Immunocytochemistry 175
6 Summary 175
Acknowledgements 178
References 178
7 Epitope mapping using synthetic peptides isi
Jane Worthington and Keith Morgan
1 Introduction 181
2 Multiple peptide synthesis on polyethylene pins 182
3 Antibody epitopes 183
General strategy 183
Membrane-bound peptides and epitope scanning 202
4 Cleavable peptides for use in identifying antibody epitopes 204
Solution-phase assay using biotinylated peptide 205
Solid-phase assay using biotinylated peptides 205
5 Synthesis of peptides to mimic epitopes of proteins for
which sequences are not available 206
Mixture strategy based on octapepudes 207
Dipeptide strategy 207
xii
Conten t s
6 Extension synthesis 207
7 The use of peptide libraries to identify antibody epitopes 207
8 T-cell epitopes 208
General strategy 208
Synthesis of cleavable peptides 210
9 Screening of cleavable peptides for the identification of
T-cell epitopes 212
Non-specific stimulation of T-cells and assay background 212
Detection systems 212
Interpretation of results 213
Further definition of epitopes 214
10 General summary and future developments 215
Acknowledgements 215
References 216
8 Epitope mapping using libraries of random
peptides displayed on phage 219
William J Dower and Steven E Cwirla
1 Introduction 219
2 Biology of the filamentous phage 220
3 An overview of the display systems 221
4 Detailed description of each phage display system 224
5 Introduction to the protocols 225
6 Constructing peptide display libraries in filamentous
phage vectors 226
7 Screening the libraries 234
Immobilization and characterization of the target antibody 234
Overview of the screening process 235
8 Characterization of the recovered clones 238
9 Concluding remarks 242
Acknowledgements 242
References 242
Appendix 1 Addresses of suppliers 245
Appendix 2 The DNA-encoded amino acids 248
Index 249
xiii
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| physical | XIX, 252 S. graph. Darst. |
| publishDate | 1994 |
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| publisher | IRL Press |
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| series | The practical approach series |
| series2 | The practical approach series |
| spelling | Peptide antigens a practical approach ed. by G. Brian Wisdom 1. ed. Oxford u.a. IRL Press 1994 XIX, 252 S. graph. Darst. txt rdacontent n rdamedia nc rdacarrier The practical approach series 144 Peptide antigens and anti-peptide antibodies are widely used in biochemistry and molecular biology for the measurement, location, and purification of specific oligopeptides. More recently the application of these reagents has expanded, for example in the identification and mapping of the binding sites of antibodies and T-cell receptors and in the identification and characterization of proteins which are known only by their primary structure. This volume provides practical guidance to the major techniques used in the exploitation of peptide antigens and anti-peptide antibodies. The chapters give detailed protocols for the prediction of epitopes, peptides synthesis, the preparation of peptide immunogens, immunoaffinity chromatography, immunoassays, and the mapping of epitopes using both synthetic peptides and phage display systems Peptide Antigens: A Practical Approach covers all practical aspects of this important and growing subject. It is a unique compendium of methods for workers in biochemistry, molecular biology, and immunology who need to use this technology in their research Antigenen gtt Antistoffen gtt Peptiden gtt Antigenic determinants Antigens Epitopes Peptides Immunology Peptides immunology Peptide (DE-588)4045125-2 gnd rswk-swf Epitop (DE-588)4209083-0 gnd rswk-swf Antigen (DE-588)4002272-9 gnd rswk-swf Immunologie (DE-588)4026637-0 gnd rswk-swf Methode (DE-588)4038971-6 gnd rswk-swf Peptide (DE-588)4045125-2 s Epitop (DE-588)4209083-0 s DE-604 Antigen (DE-588)4002272-9 s Methode (DE-588)4038971-6 s Immunologie (DE-588)4026637-0 s Wisdom, G. Sonstige oth The practical approach series 144 (DE-604)BV007921321 144 HEBIS Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=006646416&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Peptide antigens a practical approach The practical approach series Antigenen gtt Antistoffen gtt Peptiden gtt Antigenic determinants Antigens Epitopes Peptides Immunology Peptides immunology Peptide (DE-588)4045125-2 gnd Epitop (DE-588)4209083-0 gnd Antigen (DE-588)4002272-9 gnd Immunologie (DE-588)4026637-0 gnd Methode (DE-588)4038971-6 gnd |
| subject_GND | (DE-588)4045125-2 (DE-588)4209083-0 (DE-588)4002272-9 (DE-588)4026637-0 (DE-588)4038971-6 |
| title | Peptide antigens a practical approach |
| title_auth | Peptide antigens a practical approach |
| title_exact_search | Peptide antigens a practical approach |
| title_full | Peptide antigens a practical approach ed. by G. Brian Wisdom |
| title_fullStr | Peptide antigens a practical approach ed. by G. Brian Wisdom |
| title_full_unstemmed | Peptide antigens a practical approach ed. by G. Brian Wisdom |
| title_short | Peptide antigens |
| title_sort | peptide antigens a practical approach |
| title_sub | a practical approach |
| topic | Antigenen gtt Antistoffen gtt Peptiden gtt Antigenic determinants Antigens Epitopes Peptides Immunology Peptides immunology Peptide (DE-588)4045125-2 gnd Epitop (DE-588)4209083-0 gnd Antigen (DE-588)4002272-9 gnd Immunologie (DE-588)4026637-0 gnd Methode (DE-588)4038971-6 gnd |
| topic_facet | Antigenen Antistoffen Peptiden Antigenic determinants Antigens Epitopes Peptides Immunology Peptides immunology Peptide Epitop Antigen Immunologie Methode |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=006646416&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV007921321 |
| work_keys_str_mv | AT wisdomg peptideantigensapracticalapproach |